.

Friday, October 29, 2010

The optical trapping technique has been widely used in various areas to manipulate particles, cells, and so forth. The principle of trapping is based on the interaction between optical electric fields and induced linear polarizations. Here we show a novel phenomenon of trapping arising from nonlinear polarization when we trap gold nanoparticles by ultrashort near-infrared laser pulses. That is, the stable trap site is split into two equivalent positions (we call this ‘trap split’). The trap positions are aligned along the direction of the incident laser polarization. The dependencies of trap split on the trapping-laser power and wavelength were investigated. The results were successfully interpreted in terms of the nonlinear polarization caused by the femtosecond pulses. This method may give novel applications to micromachining, nanofabrication, and biological samples as well as atomic and molecular trapping at low temperatures.

Discrete dipole approximation (DDA) method is an efficient method for computing electromagnetic (EM) field of nanometer/micrometer-sized dielectric particles with arbitrary geometric shape and topology. In this work we employ the DDA method to calculate the optical force of dielectric shaped particles embedded in optical tweezers made from focused Gaussian laser beams. The EM force is calculated based on the self-consistent solution of EM field distribution and discrete dipole moment distribution within the particles. The DDA method agrees well with the Mie theory for spherical dielectric particles and this supports the effectiveness of the DDA method in handling optical forces in optical tweezers. The optical force for shaped particles such as cubes, rectangles, cylinders, and core-shell composite particles shows many interesting features. The force strongly depends on the orientation of the particle with respect to the laser beam propagation and polarization direction and the aspect ratio of the anisotropic particle. For a core-shell composite particle the zero-force balance point shifts from the particle center to its two sides. When an additional particle comes close a trapped particle, the perturbation effect strongly depends on the relative location of the center of the focused laser beam with respect to the two particles. Furthermore, the geometry of shaped particles not only affects the magnitude of the optical force but also influences the optical trap stiffness.

The compaction of DNA by the HU protein from Thermotoga maritima (TmHU) is analysed on a single-molecule level by the usage of an optical tweezers-assisted force clamp. The condensation reaction is investigated at forces between 2 and 40 pN applied to the ends of the DNA as well as in dependence on the TmHU concentration. At 2 and 5 pN, the DNA compaction down to 30% of the initial end-to-end distance takes place in two regimes. Increasing the force changes the progression of the reaction until almost nothing is observed at 40 pN. Based on the results of steered molecular dynamics simulations, the first regime of the length reduction is assigned to a primary level of DNA compaction by TmHU. The second one is supposed to correspond to the formation of higher levels of structural organisation. These findings are supported by results obtained by atomic force microscopy.

Wednesday, October 27, 2010

Vector diffraction theory is employed to investigate the focusing properties of the Gaussian beams with mixed screw and conical phase fronts. Numerical simulations show that the Gaussian beams with screw–conical phase fronts are different from both the ordinary Laguerre–Gaussian beams and the higher-order Bessel beams. Rather than forming the ring-shaped intensity distributions characteristic of optical vortices, focusing the Gaussian beams with screw–conical phase fronts produce non-symmetric spiral intensity distributions at the focal plane. The intensity distribution forms a counter-clockwise non-symmetric screw path around the focus. The rotation of intensity distributions was observed in the focal plane. The gradient force patterns of these beams focused with high NA are also investigated. The results show that the gradient force pattern shape depends principally on parameter topological charge n of the phase distribution. The gradient force pattern expands with increase in the parameter m of the phase distribution. Therefore, one can change the topological charge n or the parameter m of the phase mask to construct the tunable optical trap to meet different requirements. Its potential application might include rotational positioning of particles and accumulation of smaller non-symmetric particles towards the focus.

A new kind of hollow beams – hollow laser beams with three-dimension trap optical distribution – was put forward. With the help of the Collins formula in paraxial optical system, the analytical equation of propagation and transformation of the hollow laser beams was deduced. According to the analytical equation, the propagation properties of the kind of hollow beams that transform in free space were simulated. In the experiment, we obtained the hollow laser beams by means of the combinational optical system of reflecting positive-axis and negative-axis pyramids. The intensity of the vertical loop in different distances was tested, which shows that the analytical equation of propagation and transformation is in agreement with the result.

Focusing properties of the azimuthally polarized beam induced by a pure phase plate are investigated theoretically. The pure phase plate consists of two concentric portions, one center circle portion and one outer annular portion, through which the azimuthally polarized beam passed evolves into concentric piecewise azimuthally polarized beam. When the phase shift of the center portion is π, one ring focus may evolve into novel focal patterns with increasing radius of the center circle portion, such as cylindrical crust focus, two-ring focus, and three-ring focus. And if the geometrical parameters are unchanged, focal patterns also changes considerably with tunable phase of the center portion. Ring focus shifts along the optical axis on the increasing phase. Some optical gradient force distributions and dependence of focal shift on phase shift are also illustrated. This kind of concentric piecewise azimuthally polarized beam can be used in optical manipulation technology.

Optical forces induced by a near field are calculated for a 1-mm-sized metal particle mimicked by a jellium model and for C60 in the framework of real-time and real-space time-dependent density-functional theory combined with a nonuniform light-matter interaction formalism, fully taking account of multipole interaction. A highly localized near field nonuniformly polarizes these molecules. The locally induced polarization charges in the molecules are partly canceled by the screening charges. The polarization and screening charges generally contribute to the attractive and repulsive forces, respectively, and a sensible balance between these charges results in several peaks in the optical force as a function of the frequency of the near field. The resonance excitation does not necessarily maximally induce the net force, and the force exerted on the molecules strongly depends on the details of their electronic structures. The optical force is larger in the metal particle than in C60. We also found that the optical force depends linearly on the intensity of the near field.

Effectively organizing isolated cells to tissue elements having an appropriate microstructure is a fundamental issue in future tissue engineering, but biological cell-to-cell adhesion is too weak to assemble single cells directly. In order to overcome the difficulty, we applied an Avidin-Biotin Binding System (ABBS) to cell surfaces, and avidinylated and biotinylated cells could mutually bind in the short time they were mixed together. Unlike conventional intact cells, ABBS helped make larger spheroids. Interestingly, avidinylated and biotinylated cell adherence occurred within 1 sec using laser trapping, enabling single cell manipulation. We showed precise, direct single-cell-based tissue assembly using ABBS and optical tweezers, followed by damage-free tissue culture. The combination of ABBS and single cell manipulation has considerable potential for use in application such as tissue engineering, regenerative medicine, and drug screening system.

Jan A. van Heiningen and Reghan J. HillWe explore the design and operation of an optical-tweezers electrophoresis apparatus to resolve polymer adsorption dynamics onto a single micro-sphere in a micro-fluidic environment. Our model system represents a broader class of micro-fluidic electrophoresis experiments for biosensing and fundamental colloid and surface science diagnostics. We track the adsorption of 100 kDa poly(ethylene oxide) homopolymer onto a colloidal silica sphere that is optically trapped in a crossed parallel-plate micro-channel. The adsorption dynamics are probed on the 1 μm particle length scale with 1 s temporal resolution. Because the particle electrophoretic mobility and channel electro-osmotic flow are exquisitely sensitive to the polymer layer hydrodynamic thickness, particle dynamics can be complicated by polymer adsorption onto the micro-channel walls. Nevertheless, using experiments and a theoretical model of electro-osmotic flow in channels with non-uniform wall ζ-potentials, we show that such influences can be mitigated by adopting a symmetrical flow configuration. The equilibrium hydrodynamic layer thickness of 100 kDa poly(ethylene oxide) on colloidal silica is 10 nm at polymer concentrations 10 ppm (weight percent), with the dynamics reflecting polymer solution concentration, flow rate, and polydispersity.

Agents that alter the dynamics of hemostasis form an important part in management of conditions such as atherosclerosis, cerebrovascular disease, and bleeding diatheses. In this study, we explored the effects of heparin and tranexamic acid on the efficiency of blood coagulation. Using optical tweezers, we evaluated the pN-range micro-interaction between coagulating red blood cells (RBCs) by measuring the minimum power required to trap them. By observing the mobility of RBCs and the intensity of cellular interactions, we found that the coagulation process can be separated into three phases. The effects of heparin and tranexamic acid were examined by observing variations in cellular interaction during the coagulation phases. Heparin attenuated the interaction between RBCs and prolonged the first phase whereas the samples containing tranexamic acid bypassed the first two phases and immediately proceeded to the final one.

Tuesday, October 12, 2010

We introduce optical fiber illumination for real-time tracking of optically trapped micrometer-sized particles with microsecond time resolution. Our light source is a high-radiance mercury arc lamp and a 600μm optical fiber for short-distance illumination of the sample cell. Particle tracking is carried out with a software implemented cross-correlation algorithm following image acquisition from a CMOS camera. Our image data reveals that fiber illumination results in a signal-to-noise ratio usually one order of magnitude higher compared to standard Köhler illumination. We demonstrate position determination of a single optically trapped colloid with up to 10,000 frames per second over hours. We calibrate our optical tweezers and compare the results with quadrant photo diode measurements. Finally, we determine the positional accuracy of our setup to 2 nm by calculating the Allan variance. Our results show that neither illumination nor software algorithms limit the speed of real-time particle tracking with CMOS technology.

Laser tweezers Raman spectroscopy (LTRS) was used to acquire the Raman spectra of leukemic T lymphocytes exposed to the chemotherapy drug doxorubicin at different time points over 72 hours. Changes observed in the Raman spectra were dependent on drug exposure time and concentration. The sequence of spectral changes includes an intensity increase in lipid Raman peaks, followed by an intensity increase in DNA Raman peaks, and finally changes in DNA and protein (phenylalanine) Raman vibrations. These Raman signatures are consistent with vesicle formation, cell membrane blebbing, chromatin condensation, and the cytoplasm of dead cells during the different stages of drug-induced apoptosis. These results suggest the potential of LTRS as a real-time single cell tool for monitoring apoptosis, evaluating the efficacy of chemotherapeutic treatments, or pharmaceutical testing.

Sunday, October 10, 2010

We demonstrate negative radiation pressure on gain medium structures, such that light amplification may cause a nanoscale body to be pulled toward a light source. Optically large gain medium structures, such as slabs and spheres, as well as deep subwavelength bodies, may experience this phenomenon. The threshold gain for radiation pressure reversal is obtained analytically for Rayleigh spheres, thin cylinders, and thin slabs. This threshold vanishes when the gain medium structure is surrounded by a medium with a matched refractive index, thus eliminating the positive scattering forces.

Production of functional microtools having an arbitrary shape by self-assembly of microparticles and heat treatment above the glass transition temperature of the microparticles was developed. Polystyrene microbeads were used as a material of the microtool. A solution including microparticles was dispersed onto the silicon substrate having microtool patterns fabricated by photolithography and etching. Dispersed particles were introduced to the pattern by gravity force. Microparticles in the pattern aggregate autonomously by surface tension through evaporation of the solution. Aggregated microparticles were fused by heating above the glass transition temperature (100°C). Fused microparticles were detached from the pattern by ultrasonic treatment and used as microtools. Produced microtool has spherical part since the microtool is made of microparticles. Spherical part is suitable for trapping point of optical tweezers. We demonstrated production of microtools using self-assembly and manipulation of the fabricated microtool on a chip.

Once excess liquid gains access to airspaces of an injured lung, the act of breathing creates and destroys foam and thereby contributes to the wounding of epithelial cells by interfacial stress. Since cells are not elastic continua, but rather complex network structures composed of solid as well as liquid elements, we hypothesize that plasma membrane (PM) wounding is preceded by a phase separation, which results in blebbing. We postulate that inteventions, such as a hypertonic treatment, increase adhesive PM/cytoskeletal (CSK) interactions, thereby preventing blebbing as well as PM wounds. We formed PM tethers in alveolar epithelial cells andfibroblasts and measured their retractive force as read-out of PM/CSK adhesive interactions using optical tweezers. A 50mOsm increase in media tonicity consistently increased the tether retractive force in epithelial cells, but lowered it in fibroblasts. The osmo-response was abolished by pretreatment with Latrunculin, Cytochalasin D and calcium chelation. Epithelial cells and fibroblasts were exposed to interfacial stress in a microchannel and the fraction of wounded cells measured. Interventions which increased PM/CSK adhesive interactions prevented blebbing and were cytoprotective regardless of cell type. Finally, we exposed ex-vivo perfused rat lungs to injurious mechanical ventilation and showed that hypertonic conditioning reduced the number of wounded subpleuralalveolus resident cells to baseline levels. Our observations support the hypothesis that PM/CSK adhesive interactions are important determinants of the cells response to deforming stress and pave the way for preclinical efficacy trials of hypertonic treatment in experimental models of acute lung injury.

Counter-propagating optical traps are widely used where long working distances, axially symmetric trapping potentials, or standing light waves are required. We demonstrate that optical phase-conjugation can automatically provide a counter-propagating replica of a wide range of incident light fields in an optical trapping configuration. The resulting counter-propagating traps are self-adjusting and adapt dynamically to changes of the input light field. It is shown that not only single counter-propagating traps can be implemented by phase-conjugation, but also structured light fields can be used. This step towards more complex traps enables advanced state-of-the-art applications where multiple traps or other elaborated trapping scenarios are required. The resulting traps cannot only be used statically, but they can be rearranged in real-time and allow for interactive dynamic manipulation.

Thursday, October 7, 2010

We report the development of a multiple-trap laser tweezers Raman spectroscopy (LTRS) array for simultaneously acquiring Raman spectra of individual cells in physiological environments. This LTRS-array technique was also combined with phase contrast and fluorescence microscopy, allowing measurement of Raman spectra, refractility, and fluorescence images of individual cells with a temporal resolution of ~5 s. As a demonstration, we used this technique to monitor multiple Bacillus cereus spores germinating in a nutrient medium for up to 90min and observed the kinetics of dipicolinic acid release and uptake of nucleic acid-binding stain molecules during spore germination.

This article describes the design, implementation and characterization of a novel optical tweezer system. The system utilizes a deformable mirror, wavefront sensor and controller to manipulate an optically trapped micro-particle within a small chamber. This method for optical trapping employs technology adopted from astronomical instrumentation; in particular, adaptive optics. A deformable mirror is employed to control the wavefront phase of a laser beam before it is imaged into a chamber by a high numerical aperture microscope objective lens. The wavefront phase is measured by a Shack-Hartman wavefront sensor and the particle's position monitored by a video camera. The goals of the work presented here are to trap particles ranging in size from 1 μm to 10 μm; create a suitable controller for moving trapped particles in three dimensions; image the trapped particle; determine the prototype system's performance specifications; and determine the trap stiffness.

It is well known that optical force fields are not conservative. This has important consequences for the thermal motion of optically trapped dielectric spheres. In particular, the spheres do not reach thermodynamic equilibrium. Instead, a steady state is achieved in which the stochastic trajectory contains an underlying deterministic bias toward cyclic motion, and the energy of the sphere deviates from that implied by the equipartition theorem. Such effects are second order and only observed at low trap powers when the sphere is able to explore regions of the trap beyond the linear regime. Analogous effects may be expected for particles of less than spherical symmetry. However, in this case the effects are first order and depend on the linear term in the optical force field. As such they are not suppressed by increases in beam power, although the frequency and amplitude of the cyclic motion will be affected by it. In this paper, we present an analysis of the first-order nonconservative behavior of nonspherical particles in optical traps. The analysis is supported by optical force calculations and Brownian dynamics simulations of dielectric microrods held vertically in Gaussian optical traps.

Near-infrared laser (785-nm)-excited Raman spectra from a red blood cell, optically trapped using the same laser beam, show significant changes as a function of trapping duration even at trapping power level of a few milliwatts. These changes in the Raman spectra and the bright-field images of the trapped cell, which show a gradual accumulation of the cell mass at the trap focus, suggest photoinduced aggregation of intracellular heme. The possible role of photoinduced protein denaturation and hemichrome formation in the observed aggregation of heme is discussed.

The regulation of actin filament networks by various proteins has essential roles in the growth cone dynamics. In this study we focused on the actin–myosininteraction which has been suggested to be an important player in the neurite extension. We examined in vitro how the decoration of actin filaments with a side-binding protein, drebrin-E, affects the motile properties of an intracellular transporter myosin V. Single myosin V molecules landed on the drebrin-E-decorated actin filaments with a lower frequency and ran over shorter distances; however, their velocities were normal. Furthermore, the analysis of the movement of myosin V molecules in the optical trap revealed that the decoration of actin filaments with drebrin-E markedly increased the load-sensitivity of the myosin V stepping. These results are attributable to the delay in the attachment of the motor’s leading head (ADP·Pi state) to actin, induced by the competitive binding of drebrin-E to actin, whereas the rate of ADP release from the trailing head (the rate-limiting step in the ATPase cycle of myosin V) is unaffected. Our study indicates that, in addition to the regulation of binding affinity of myosin V, drebrin-E also modulates the chemo-mechanical coupling in the motile myosin V molecules, presumably affecting the movement of the growth cone.

In this paper, we investigate an optical-trap-based method for the detection of structural changes of the red blood cell (RBC) membrane affected by$hbox{Ca}^{2+}$ ions. Individual cells are immobilized by the use of optical tweezers and are monitored live, while the concentration of $hbox{Ca}^{2+}$ions in the buffer is changed simultaneously. $hbox{Ca}^{2+}$ ions are known to affect the cells' membrane morphology. These changes are attributed to the formation of calcium-induced hydrophobic aggregates of phospholipid molecules in the RBC membrane, resulting in a net change in membrane rigidity. Membrane deformation results in the change of effective radius and the drag coefficient of the cell, both of which affect the Brownian motion of the cell in solution. This motion is indirectly measurable by monitoring the forward scattering light and its dependence on the size and drag coefficient of the cell. We show the relationship between the $hbox{Ca}^{2+}$ ion concentration and the optical trap specifications. The results are in agreement with previous biological studies and the phase contrast observations of living RBCs under investigation.

A high-precision study of hindered diffusion of a sphere due to its proximity to a solid interface was performed using an optical tweezer combined with digital holography microscopy and a phase-sensitive detection technique. The study provides a confirmation, with high accuracy and no adjustable parameters, of Faxen’s law which describes the variation of the diffusivity of a sphere as a function of its distance from the wall in a Newtonian liquid due to hydrodynamic effects. This general technique is useful for application in microfluidics and lubrication of small devices with moving parts.

Radially polarized beams, focused by a high numerical aperture (NA) objective, have non-propagating fields along the propagation axis in the focal region, which leads to a higher axial trapping efficiency in comparison with linearly polarized beams. We propose a design for converting a lowest-order radially polarized beam (RTEM01) to a double-ring radial polarization distribution (DR R-TEM01) through a specially designed polarization rotator. The phases of the two rings of this beam differ by pi. Numerical results evaluated by rigorous T-matrix method show that the DR R-TEM01 beam can improve the axial trapping efficiency compared with the R-TEM01 beam, provided that the size of trapped particles is of order of the wavelength of the beam.

The vapor pressures of two dicarboxylic acids, malonic acid and glutaric acid, are determined by the measurement of the evaporation rate of the dicarboxylic acids from single levitated particles. Two laboratory methods were used to isolate single particles, an electrodynamic balance and optical tweezers (glutaric acid only). The declining sizes of individual aerosol particles over time were followed using elastic Mie scattering or cavity enhanced Raman scattering. Experiments were conducted over the temperature range of 280-304 K and a range of relative humidities. The subcooled liquid vapor pressures of malonic and glutaric acid at 298.15 K were found to be 6.7-1.2+2.6 ×10-4 and 11.2-4.7+9.6 ×10-4 Pa, respectively, and the standard enthalpies of vaporization were respectively 141.9 ± 19.9 and 100.8 ± 23.9 kJ mol-1. The vapor pressures of both glutaric acid and malonic acid in single particles composed of mixed inorganic/organic composition were found to be independent of salt concentration within the uncertainty of the measurements. Results are compared with previous laboratory determinations and theoretical predictions.